1. Field of the Invention
The present invention relates to novel compounds that function as enzyme inhibitors, and particularly to a new class of non-peptidic inhibitors of proteolytic enzymes.
2. Related Art
Proteases are enzymes that cleave proteins at single, specific peptide bonds. Proteases can beclassified into four generic classes: serine, thiol or cysteinyl, acid or aspartyl, and metalloproteases (Cuypers et al., J. Biol. Chem. 257:7086 (1982)). Proteases are essential to a variety of biological activities, such as digestion, formation and dissolution of blood clots, reproduction and the immune reaction to foreign cells and organisms. Aberrant proteolysis is associated with a number of disease states in man and other mammals. The human neutrophil proteases, elastase and cathepsin G, have been implicated as contributing to disease states marked by tissue destruction. These disease states include emphysema, rheumatoid arthritis, comeal ulcers and glomerular nephritis. (Barret, in Enzyme Inhibitors as Drugs, Sandler, ed., University Park Press, Baltimore, (1980)). Additional proteases such as plasmin, C-1 esterase, C-3 convertase, urokinase, plasminogen activator, acrosin, and kallikreins play key roles in normal biological functions of mammals. In many instances, it is beneficial to disrupt the function of one or more proteolytic enzymes in the course of therapeutically treating a mammal.
Serine proteases include such enzymes as elastase (human leukocyte), cathepsin G, plasmin, C-1 esterase, C-3 convertase, urokinase, plasminogen activator, acrosin, chymotrypsin, trypsin, thrombin, factor Xa and kallikreins.
Human leukocyte elastase is released by polymorphonuclear leukocytes at sites of inflammation and thus is a contributing cause for a number of disease states. Cathepsin G is another human neutrophil serine protease. Compounds with the ability to inhibit the activity of these enzymes are expected to have an anti-inflammatory effect useful in the treatment of gout, rheumatoid arthritis and other inflammatory diseases, and in the treatment of emphysema Chymotrypsin and trypsin are digestive enzymes. Inhibitors of these enzymes are useful in treating pancreatitis. Inhibitors of urokinase and plasminogen activator are useful in treating excessive cell growth disease states, such as benign prostatic hypertrophy, prostatic carcinoma and psoriasis.
The serine protease thrombin occupies a central role in hemostasis and thrombosis, and as a multifactorial protein, induces a number of effects on platelets, endothelial cells, smooth muscle cells, leukocytes, the heart, and neurons (Tapparelli et al., Trends in Pharmacological Sciences 14:366-376 (1993); Lefkovits and Topol, Circulation 90(3):1522-1536 (1994); Harker, Blood Coagulation and Fibrinolysis 5 (Suppl 1):S47-S58 (1994)). Activation of the coagulation cascade through either the intrinsic pathway (contact activation) or the extrinsic pathway (activation by exposure of plasma to a non-endothelial surface, damage to vessel walls or tissue factor release) leads to a series of biochemical events that converge on thrombin. Thrombin cleaves fibrinogen ultimately leading to a hemostatic plug (clot formation), potently activates platelets through a unique proteolytic cleavage of the cell surface thrombin receptor (Coughlin, Seminars in Hematology 31(4):270-277 (1994)), and autoamplifies its own production through a feedback mechanism. Thus, inhibitors of thrombin function have therapeutic potential in a host of cardiovascular and non-cardiovascular diseases, including: myocardial infarction; unstable angina; stroke; restenosis; deep vein thrombosis; disseminated intravascular coagulation caused by trauma, sepsis or tumor metastasis;
hemodialysis; cardiopulmonary bypass surgery; adult respiratory distress syndrome; endotoxic shock; rheumatoid arthritis; ulcerative colitis; induration; metastasis; hypercoagulability during chemotherapy; Alzheimer""s disease; and Down""s syndrome.
Factor Xa is another serine protease in the coagulation pathway. Factor Xa associates with factor Va and calcium on a phospholipid membrane thereby forming a prothrombinase complex. This prothrombinase complex then converts prothrombin to thrombin (Claeson, Blood Coagulation and Fibrinolysis 5:411-436 (1994); Harker, Blood Coagulation and Fibrinolysis 5 (Suppl 1):S47-S58 (1994)). Inhibitors of factor Xa are thought to offer an advantage over agents that directly inhibit thrombin since direct thrombin inhibitors still permit significant new thrombin generation (Lefkovits and Topol, Circulation 90(3):1522-1536 (1994); Harker, Blood Coagulation and Fibrinolysis S (Suppl 1):S47-S58 (1994)).
A need continues to exist for non-peptidic compounds that are potent and selective protease inhibitors, and which possess greater bioavailability and fewer side-effects than currently available protease inhibitors. Accordingly, new classes of potent protease inhibitors, characterized by potent inhibitory capacity and low mammalian toxicity, are potentially valuable therapeutic agents for a variety of conditions, including treatment of a number of mammalian proteolytic disease states.
The present invention is directed to novel compounds having one of Formulae I-III (below). Also provided are processes for preparing compounds of Formulae I-III. The novel compounds of the present invention are potent inhibitors of proteases, especially trypsin-like serine proteases, such as chymotrypsin, trypsin, thrombin, plasmin and factor Xa. Certain of the compounds exhibit antithrombotic activity via direct inhibition of thrombin, or are intermediates useful for forming compounds having antithrombotic activity. Other compounds are inhibitors of trypsin and/or chymotrypsin, and are therefore useful in treating pancreatitis. Also provided are methods of inhibiting or treating aberrant proteolysis in a mammal and methods of treating thrombosis, ischemia, stroke, restenosis or inflammation in a mammal by administering an effective amount of a compound of Formulae I-III. Further provided are pharmaceutical compositions comprising a compound of Formulae I-III and one or more pharmaceutically acceptable carriers or diluents.
Compounds of the present invention include compounds having one of Formulae I-III: 
or solvates, hydrates or pharmaceutically acceptable salts thereof; wherein:
Z is one of xe2x80x94NR10SO2xe2x80x94, xe2x80x94SO2NR10xe2x80x94, xe2x80x94NR10C(RyRz)xe2x80x94, xe2x80x94C(RyRz)NR10xe2x80x94, xe2x80x94OSO2xe2x80x94, xe2x80x94SO2Oxe2x80x94, xe2x80x94OC(RyRz)xe2x80x94, xe2x80x94(RyRz)Oxe2x80x94, xe2x80x94NR10COxe2x80x94 or xe2x80x94CONR10xe2x80x94;
Ry and Rz are each independently one of hydrogen, alkyl, cycloalkyl, aryl, aralkyl, hydroxyalkyl, carboxyalkyl, aminoalkyl, monoalkylaminoalkyl, dialkylaminoalkyl or carboxy;
R1 is one of allyl, cycloalkyl, alkenyl, alkynyl, aryl, araLkyl or heteroaryl, any of which may be optionally substituted;
R2, R3 and R4 are each independently one of hydrogen, alkyl, cycloalkyl, alkenyl, alkynyl, aryl, aralkyl, heteroaryl, trifluoromethyl, halogen, hydroxyalkyl, cyano, nitro, carboxamide, xe2x80x94CO2Rx, xe2x80x94CH2ORx or xe2x80x94ORx, or when present on adjacent carbon atoms, R2 and R3 may also be taken together to form one of xe2x80x94CHxe2x95x90CHxe2x80x94CHxe2x95x90CHxe2x80x94 or xe2x80x94(CH2)qxe2x80x94, where q is from 2 to 6, and R4 is defined as above;
Rx, in each instance, is independently one of hydrogen, alkyl or cycloalkyl wherein said alkyl or cycloalkyl groups may optionally have one or more unsaturations;
Y is one ofxe2x80x94Oxe2x80x94, xe2x80x94NR10xe2x80x94, xe2x80x94Sxe2x80x94, xe2x80x94CHR10xe2x80x94 or a covalent bond;
W is N or CR10;
R5 is one of hydrogen, alkyl, aralkyl, aryl, hydroxyalkyl or carboxyalkyl;
R6, in each instance, is independently one of hydrogen, alkyl, hydroxy, alkoxy, aryloxy, aralkoxy, alkoxycarbonyloxy, cyano or xe2x80x94CO2Rw, where Rw is alkyl or cycloalkyl;
R7 and R8 are each independently one of hydrogen, alkyl, aralkyl, aryl, hydroxyalkyl or carboxyalkyl, or R7 and R8 are taken together to form xe2x80x94(CH2)yxe2x80x94, where y is zero, 1 or 2, with the proviso that when W is N, y cannot be zero or 1;
R9 is one of hydrogen, alkyl, cycloalkyl or aryl, wherein said alkyl, cycloalkyl or aryl can be optionally substituted with amino, monoalkylamino, dialkylamino, alkoxy, hydroxy, carboxy, alkoxycarbonyl, aryloxycarbonyl, aralkoxycarbonyl, aryl, heteroaryl, acylamino, cyano or trifluoromethyl;
R10, in each instance, is independently one of hydrogen, alkyl, aralkyl, aryl, hydroxyalkyl, aminoalkyl, monoalkylamino(C2-10)alkyl, dialkylamino (C2-10)alkyl or carboxyalkyl;
Rxe2x80x2 is one of hydrogen, alkyl, cycloalkyl, aryl, aralkyl, heteroaryl, trifluoromethyl, halogen, hydroxyalkyl, cyano, nitro, carboxamide, carboxy, alkoxycarbonyl or alkoxyalkyl;
n is from zero to 8, with the proviso that when W is N and Y is other than xe2x80x94CHR10xe2x80x94, then n is from 2 to 8; and
m is from 1 to 4, provided that when W is N, then m is not 1.
A preferred group of compounds falling within the scope of the present invention include compounds of Formulae I-III wherein:
Z is one ofxe2x80x94SO2Oxe2x80x94, xe2x80x94SO2NR10xe2x80x94, xe2x80x94C(RyRz)Oxe2x80x94 or xe2x80x94OC(RyRz)xe2x80x94, where Ry and Rz are each hydrogen;
R1 is one of C6-10 aryl, pyridinyl, quinizolinyl, quinolinyl or tetrahydroquinolinyl, any of which is optionally substituted by one or two of hydroxy, nitro, trifluoromethyl, halogen, C1-6 alkyl, C1-6 alkoxy, C1-6 aminoalkyl, C1-6 aminoalkoxy, amino, mono(C1-4)alkylamino, di(C1-4)alkylamino, C2-6 alkoxycarbonylamino, C2-6 alkoxycarbonyl, carboxy, C1-6 hydroxyalkyl, C2-6 hydroxyalkoxy, C2-10 mono(carboxyalkyl)amino, di(C2-10 carboxyalkyl)amino, C6-14 ar(C1-6) alkoxycarbonyl, C2-6 alkynylcarbonyl, C1-6 alkylsulfonyl, C2-6 alkenylsulfonyl, C2-6 alkynylsulfonyl, C1-6 alkylsulfinyl, C1-6 alkylsulfonamido, amidino, guanidino, C1-6 alkyliminoamino, formyliminoamino, C2-6 carboxyalkoxy, C2-6 carboxyalkyl, carboxyalkylamino, cyano, trifluoromethoxy, and perfluoroethoxy;
R2, R3 and R4 are independently one of hydrogen, C1-6 alkyl, C3-8 cycloalkyl, phenyl, benzyl, trifluoromethyl, halogen, hydroxy(C1-8)alkyl, cyano, nitro, carboxamide, carboxy, C1-4 alkoxycarbonyl, C1-4 alkoxymethyl or C1-4 alkoxy; or alternatively, R2 and R3, when present on adjacent carbon atoms, may also be taken together to form one of xe2x80x94CHxe2x95x90CHxe2x80x94CHxe2x95x90CHxe2x80x94 or xe2x80x94CH2)qxe2x80x94, where q is from 2 to 6, and R4 is as defined above;
Y is one of xe2x80x94Oxe2x80x94, xe2x80x94Sxe2x80x94, xe2x80x94NR10xe2x80x94, or a covalent bond;
W is N or CR10;
R5 is one of hydrogen, C1-4 alkyl, C2-10 carboxyalkyl or C2-10 hydroxyalkyl;
R6, in each instance, is one of hydrogen, C1-4 alkyl, hydroxy, C1-4 alkoxy, phenoxy, C1-4 alkyloxycarbonyl or cyano;
R7 and R8 are independently one of hydrogen, C1-6 alkyl, C2-10 carboxyalkyl or C2-10 hydroxyalkyl, or R7 and R8 are taken together to form xe2x80x94CH2)yxe2x80x94 where y is 0, 1 or 2, provided that when W is N, y cannot be 0 or 1;
R9 is hydrogen; or C1-10 alkyl, optionally substituted with amino, mono(C1-4)alkylamino, C1-6 alkoxy, hydroxy, carboxy, phenyl, alkyloxycarbonyl, aralkoxycarbonyl, C1-6 acylamino, cyano or trifluoromethyl;
R10, in each instance, is independently hydrogen, C1-6 alkyl, benzyl, phenyl, C2-10 hydroxyalkyl, C2-10 aminoalkyl, C1-4 monoalylamino(C2-8)alkyl, C1-4 dialkylamino(C2-8)alkyl or C2-10 carboxyalkyl;
Rxe2x80x2 is one of hydrogen, C1-6 alkyl, C3-8, cycloalkyl, phenyl, benzyl, trifluoromethyl, halogen, hydroxy(C1-8)alkyl, cyano, nitro, carboxamide, carboxy, alkoxycarbonyl, alkoxymethyl or alkoxy;
n is from zero to 8, with the proviso that when W is N, then n is from 2 to 8; and
m is from 1 to 4, provided that when W is N, then m is not 1.
An especially preferred group of compounds include compounds of Formulae I-III wherein:
Z is one ofxe2x80x94SO2Oxe2x80x94, xe2x80x94SO2NR10xe2x80x94, xe2x80x94CH2Oxe2x80x94 or xe2x80x94OCH2xe2x80x94;
R1 is one of phenyl or naphthyl, optionally substituted by one or two of chloro or dimethylamino;
R2 and R3 are each hydrogen or R2 and R3 may also be taken together to form xe2x80x94CHxe2x95x90CHxe2x80x94CHxe2x95x90CHxe2x80x94;
R4 is one of hydrogen, methyl, methoxy or trifluoromethyl;
Y is one of O or NR10;
W is N or CR10;
R5 is one of hydrogen, C1-6 alkyl, C2-10 hydroxyalkyl or C2-10 carboxyalkyl;
R6, in each instance is hydrogen or hydroxy;
R7 and R 8 are independently one of hydrogen, C1-6 alkyl, C2-10 hydroxyalkyl or C2-10 carboxyalkyl, or R7 and R8 are taken together to form xe2x80x94(CH2)yxe2x80x94, where y is zero, 1 or 2, with the proviso that when W is N, y cannot be zero or 1;
R9 is hydrogen or C1-4 alkyl;
R10, in each instance, is independently hydrogen, C1-4 alkyl, C2-4 hydroxyalkyl, C2-4 carboxyalkyl, C2-4 aminoalkyl, dimethylamino(C2-8)alkyl, methylamino(C2-8)alkyl;
Rxe2x80x2 is hydrogen, methyl, methoxy or trifluoromethyl;
n is from zero to 4, with the proviso that when W is N, then n is 2 to 4; and m is 1, 2 or 3.
Useful compounds falling within the scope of Formula I include compounds having one of Formulae IV-VI: 
or solvates, hydrates or pharmaceutically acceptable salts thereof; wherein:
Z, R1, R2, R3, R4, Y, R6, R9 and R10 are defined as above for Formulae I-III;
R18 is one of hydrogen, alkyl, aralkyl, aryl, C2-10 hydroxyalkyl or C2-10 carboxyalkyl;
a is from 1 to 8, provided that when Y is other than xe2x80x94CHR10xe2x80x94, then a is from 2 to 8;
b is from 1 to 8; and
c is from 1 to 13, provided that when Y is other than xe2x80x94CHR10xe2x80x94, then c is from 2-13.
Preferred compounds falling within the scope of Formula II include compounds having one of Formulae VII-IX: 
or solvates, hydrates or pharmaceutically acceptable salts thereof; wherein:
Z, R1, R2, R3, R4, Y, R6, R9 and R10 are defined as above for Formulae I-III;
R18 is one of hydrogen, alkyl, aralkyl, aryl, C2-10 hydroxyalkyl or C2-10 carboxyalkyl;
a is from 1 to 8, provided that when Y is other than xe2x80x94CHR10xe2x80x94, then a is from 2 to 8;
b is from 1 to 8; and
c is from 1 to 13, provided that when Y is other than xe2x80x94CHR10xe2x80x94, then c is from 2-13.
Preferred compounds falling within the scope of Formula III include compounds having one of Formulae X or XI: 
or solvates, hydrates or pharmaceutically acceptable salts thereof, wherein:
Z, R1, R2, R3, R4, Y, R6, R9 and R10 are defmed as above for Formulae I-III;
R18 is one of alkyl, aralkyl, aryl, C2-10 hydroxyalkyl or C2-10 carboxyalkyl;
d is from 1 to 8; and
e is from 1 to 8.
The moiety xe2x80x94Zxe2x80x94R1 of Formulae I-XI is attached to the benzene ring in a position ortho-, meta- or para- to Y.
The amidino moiety (xe2x80x94C(xe2x95x90NR6)NR6R6) of Formulae III, X and XI can be attached in the ortho-, meta- or para- positions.
Preferred compounds of the present invention are those of Formula I-XI wherein Y is one of divalent oxygen (xe2x80x94Oxe2x80x94) or xe2x80x94NR10xe2x80x94 and Z is one of xe2x80x94SO2NR10xe2x80x94, xe2x80x94SO2Oxe2x80x94 or xe2x80x94CH2Oxe2x80x94.
Preferred compounds of the present invention are those of Formula I-XI wherein R1 is one of C1-12 alkyl, C4-7 cycloalkyl, C2-8 alkenyl, C2-8 alkynyl or C6-14 aryl, especially C6-10 aryl, any of which is optionally substituted. Substituents that can be optionally present on the RI moieties include one or more, preferably one or two, hydroxy, nitro, trifluoromethyl, halogen, alkoxy, aminoalkoxy, aminoalkyl, hydroxyalkyl, hydroxyalkoxy, cyano, amino, monoalkylamino, dialkylamino, carboxy, carboxyalkyl, carboxyalkoxy, mono(hydroxyalkyl)amino, di(hydroxyalkyl)amino, mono(carboxyalkyl)amino, di(carboxyalkyl)amino, alkoxycarbonylamino, alkoxycarbonyl, aralkoxycarbonyl, alkenylcarbonyl, alkynylcarbonyl, alkylsulfonyl, alkenylsulfonyl, alkynylsulfonyl, alkylsulfinyl, alkylsulfonamido, arnidino, guanidino, alkyliminoamino, formyliminoamino, trifluoromethoxy or perfluoroethoxy. A further substituent on aryl, cycloalkyl, alkenyl, alkynyl and aralkyl moities of RI includes one or more, preferably one or two, alkyl moieties. Preferred values of optional substituents on R1 include hydroxy, nitro, trifluoromethyl, halogen, C1-6 alkyl, C1-6 alkoxy, C1-6 aminoalkyl, C1-6 aminoalkoxy, amino, mono(C1-4)alkylamino, di(C1-4)alkylamino, C2-6 alkoxycarbonylamino, C2-6 alkoxycarbonyl, carboxy, C1-6 hydroxyalkyl, C2-10 mono(carboxyalkyl)amino, di(C2-10 carboxyalkyl)amino, C6-14 ar(C1-6 alkoxycarbonyl, C2-6 alkynylcarbonyl, C1-6 alkylsulfonyl, C2-6 alkenylsulfonyl, C2-6 alkynylsulfonyl, C1-6 alkylsulfinyl, C1-6 alkylsulfonamido, amidino, guanidino, C1-6 alkyliminoamino, formyliminoamino, C2-6 carboxyalkoxy, carboxyalkylamino, cyano, trifluoromethoxy, and perfluoroethoxy.
An additional preferred group of compounds are those compounds of Formulae I-XI wherein R1 is heteroaryl or substituted heteroaryl. Preferred R1 heteroaryl groups include pyridyl, thienyl, chromenyl, benzoxazolyl, quinazolinyl, quinolinyl and tetrahydroquinolinyl, with pyridyl, quinazolinyl, quinolinyl and tetrahydroquinolinyl being most preferred. Preferred compounds when R1 is substituted heteroaryl include those compounds having one of the heteroaryl groups mentioned as preferred that have one or more, preferably one or two, substituents that are listed in the preceding paragraph.
Useful values of R1 include phenyl, chlorophenyl, iodophenyl, dichlorophenyl, bromophenyl, trifluoromethylphenyl, di(trifluoromethyl)phenyl, methylphenyl, t-butylphenyl, methoxyphenyl, dimethoxyphenyl, hydroxyphenyl, carboxyphenyl, aminophenyl, methylaminophenyl, n-butylaminophenyl, amidinophenyl, guanidinophenyl, formyliminoaminophenyl, acetimidoylaminophenyl, methoxycarbonylphenyl, ethoxycarbonylphenyl, carboxymethoxyphenyl, naphthyl, hydroxynaphthyl, cyclohexyl, cyclopentyl, 2-propylbutyl, quinolinyl and tetrahydroquinolinyl.
The groups R2, R3 and R4 in Formulae I-XI substitute for any remaining hydrogen atoms on the benzene ring after allowing for attachment of the moiety xe2x80x94Zxe2x80x94R1. Preferred compounds are those where R2, R3 and R4 are independently hydrogen, C1-4 alkyl, C4-7 cycloalkyl, C6-14 aryl, especially C6-10 aryl, C6-10 ar(C1-4)alkyl, trifluoromethyl, halogen, hydroxyalkyl, cyano, nitro, carboxamide, carboxy, alkoxycarbonyl, carboxymethyl, alkoxycarbonylmethyl, or cycloalkyloxycarbonyl. Alternatively, R2 and R3, when attached to adjacent carbon atoms on the benzene ring, are one of xe2x80x94CHxe2x95x90CHxe2x80x94CHxe2x95x90CHxe2x80x94 or xe2x80x94(CH2)qxe2x80x94, where q is from 2 to 6, thereby forming a fused ring. Preferred values of R2 together with R3 include xe2x80x94CHxe2x95x90CHxe2x80x94CHxe2x95x90CHxe2x80x94, xe2x80x94CH2xe2x80x94CH2xe2x80x94CH2xe2x80x94 and xe2x80x94CH2xe2x80x94CH2xe2x80x94CH2xe2x80x94CH2. When R2 and R3 together form a fused ring, R4 is preferably hydrogen.
Useful values of R2, R3 and R4 include hydrogen, methyl, ethyl, chloro, bromo, trifluoromethyl, hydroxymethyl, methoxy, ethoxy, carboxamide, nitro, phenyl, cyclopropyl, hydroxy, isopropyl, methoxycarbonyl, ethoxycarbonyl and benzyl. Useful values of R2, R3 and R4 also include R2 and R3 together forming xe2x80x94CHxe2x95x90CHxe2x80x94CHxe2x95x90CH or xe2x80x94CH2xe2x80x94CH2xe2x80x94CH2xe2x80x94 and R4 being hydrogen.
Preferred values of R6 in Formulae I-XI are hydrogen, hydroxy, C1-6alkyl, C1-6 alkoxy, cyano or xe2x80x94CO2Rw, where Rw, in each instance, is preferably one of C1-4 alkyl or C4-7 cycloalkyl. Suitable values of R6 include hydrogen, methyl, ethyl, propyl, n-butyl, hydroxy, methoxy, ethoxy, cyano, xe2x80x94CO2CH3, xe2x80x94CO2CH2CH3 and xe2x80x94CO2CH2CH2CH3. In the most preferred embodiments, each R6 is hydrogen.
Preferred compounds include compounds of Formulae I and II, where R7 and R8 are independently one of hydrogen, C1-6 alkyl, C6-10 ar(C1-6)alkyl, C6-10 aryl, C2-10 hydroxyalkyl or C2-7 carboxyalkyl, or R7 and R8 are taken together to form xe2x80x94CH2)yxe2x80x94, where y is most preferably 2. Useful values of R7 and R8 include hydrogen, methyl, ethyl, propyl, n-butyl, benzyl, phenylethyl, 2-hydroxyethyl, 3-hydroxypropyl, 4-hydroxybutyl, 2-carboxymethyl, 3-carboxyethyl and 4-carboxypropyl.
Preferred compounds are those of Formulae I, IV, V and VI, wherein R9 is C1-10 hydrogen or alkyl optionally substituted by one, two or three of, preferably one of, amino, monoalkylamino, dialkylamino, alkoxy, hydroxy, alkoxycarbonyl, aryloxycarbonly, aralkoxycarbonyl, carboalkoxy, phenyl, cyano, trifluoromethyl, acetylamino, pyridyl, thienyl, furyl, pyrrolyl or imidazolyl.
Suitable values of R9 include hydrogen, methyl, ethyl, propyl, n-butyl, benzyl, phenethyl, 2-hydroxyethyl, 3-hydroxypropyl, 4-hydroxybutyl, carboxymethyl and carboxyethyl.
Preferred values of R10 in Formulae I-XI include hydrogen, C1-6 alkyl, C6-10 ar(C1-6)alkyl, C6-10 aryl, C2-10 hydroxyalkyl C2-10 aminoalkyl, C2-7 carboxyalkyl, mono(C1-4 alkyl)amino(C1-8)alkyl, and di(C1-4 alkyl)amino (C1-8)alkyl. Suitable values of R10 include methyl, ethyl, propyl, n-butyl, benzyl, phenylethyl, 2-hydroxyethyl, 3-hydroxypropyl, 4-hydroxybutyl, 2-aminoethyl, carboxymethyl, 2-carboxyethyl, 3carboxypropyl and 2-(dimethylamino)ethyl.
Preferred values of n in Formulae I-III include from 1 to 6, more preferably from 1 to 4, and most preferably 1 or 2, with the proviso that when W is N and Y is other than xe2x80x94CHR10xe2x80x94, then n is not 1. Preferred values of m include from 1 to 4, more preferably 1, 2 or 3, provided that when W is N, then m is not 1.
Preferred values of R5 in Formula III include is one of hydrogen, C1-4 alkyl, phenyl, benzyl, phenethyl, C2-10 carboxyalkyl and C2-10 hydroxyalkyl. Especially preferred values are hydrogen, C1-6 alkyl, C2-10 hydroxyalkyl and C2-10 carboxyalkyl. Suitable values of R5 include hydrogen, methyl, hydroxymethyl, hydroxyethyl, carboxymethyl and carboxyethyl.
Preferred values of Rxe2x80x2 in Formula III include hydrogen, C1-6 alkyl, C3-8 cycloalkyl, phenyl, benzyl, trifluoromethyl, halogen, hydroxy(C1-8)alkyl, cyano, nitro, carboxamide, carboxy, alkoxycarbonyl, alkoxymethyl and alkoxy. Suitable values of Rxe2x80x2 include hydrogen, methyl, methoxy and trifluoromethyl;
Preferred values of xe2x80x9caxe2x80x9d in Formulae IV and VII include from 1 to 6, more preferably from 1 to 4, and most preferably 1 or 2, with the proviso that when Y is other than xe2x80x94CHR10xe2x80x94, then n is not 1.
Preferred values of xe2x80x9cbxe2x80x9d in Formulae V and VIII include from 1 to 6, preferably from 1 to 4, and most preferably 1 or 2.
Preferred values of xe2x80x9ccxe2x80x9d in Formulae VI and IX include from 1 to 8, more preferably from 1 to 6, and most preferably 1, 2, 3, or 4.
Preferred values of xe2x80x9cdxe2x80x9d and xe2x80x9cexe2x80x9d in Fromulae V and XI include from 1 to 6, preferably from 1 to 4, and most preferably 1 or 2.
Preferred compounds of Formulae VI, IX and XI are those where R18 is independently one of hydrogen, C1-6 alkyl, C6-10 ar(C1-6)alkyl, C6-10 aryl, C2-10 hydroxyallyl and C2-7 carboxyalkyl. Useful values of R18 include hydrogen, methyl, ethyl, propyl, n-butyl, benzyl, phenylethyl, 2-hydroxyethyl, 3-hydroxypropyl, 4-hydroxybutyl, 2-carboxymethyl, 3-carboxyethyl and 4-carboxypropyl. Most preferred compounds are those where R18 is hydrogen.
Specific compounds within the scope of the invention include the following examples:
2-chlorobenzenesulfonic acid 3-[(1-acetimidoylpiperidin-4-yl)methoxy]-5-methylphenyl ester hydrochloride;
3-(2-chlorobenzyloxy)-5-methyl-1- [2-(1 -acetimidoyl)piperazin-4-yl]]ethoxybenzene diacetic acid salt;
N-[2-(N,N-dimethylamino)ethyl]-N-[2-[[4-(1-acetimidoyl)amino]butoxy]-4-methylphenyl]benzenesulfonamide dihydrochloride;
N-benzyl-N- [[[3 -(1-acetimidoyl)piperidin-4-yl]methylamino]phenyl]-benzenesulfonamide; 3-chlorobenzenesulfonic acid 3-[[(1-acetimidoyl)piperidin-4-yl]methoxy]-5-methylphenyl ester hydrochloride;
2-chlorobenzenesulfonic acid 3-[(3-amidinophenyl)methoxy]-5-methylphenyl ester hydrochloride;
2-chlorobenzenesulfonic acid 3-[[3-(N-hydroxy)amidinophenyl]methoxy]-5-methylphenyl ester hydrochloride;
2,3-dichlorobenzenesulfonic acid 3-[[(l-acetimidoyl)piperidin-4-yl]methoxy]-5-methylphenyl ester hydrochloride;
2-chloro-N-[[3 - [(1-acetimidoyl)piperidin-4-yl]methoxy] -5-trifluoromethylphenyl]benzenesulfonamide hydrochloride;
2-chloro-N-(5-carboxypentyl)-N-[[3-[(1 -acetimidoyl)piperidin-4-yl]methoxy]-5-trifluoromethylphenyl]benzenesulfonamide;
1-(5-N,N-dimethylamino)naphthalenesulfonic acid 3-[[(1-acetimidoyl)piperidin-3-yl]methoxy]-5-methoxyphenyl ester hydrochloride;
2-chlorobenzenesulfonic acid 1-[[(1-acetimidoyl)piperidin-4-yl]methoxy]naphthalen-3-yl ester acetic acid salt;
3-[(2-chlorophenoxy)methyl]-[[(1-acetimidoyl)piperidin-4-yl]methoxy]benzene acetic acid salt;
2-Chlorobenzenesulfonic acid 3-[(4-amidinophenyl)methoxy]-5-methylphenyl ester hydrochloride;
2-chiorobenzenesulfonic acid 3-[(3-amidinophenyl)methoxy]phenyl ester hydrochloride;
2-chlorobenzenesulfonic acid 3-[5-amidinopentyloxy]-5-methylphenyl ester acetic acid salt;
2-chlorobenzenesulfonic acid 3-[3-amidinopropoxy]-5-methylphenyl ester hydrochloride; and
2-chlorobenzenesulfonic acid 3-[[3-(N-methylamidino)phenyl]methoxy]-5-methylphenyl ester hydrochloride.
It is also to be understood that the present invention is considered to include stereoisomers as well as optical isomers, e.g. mixtures of enantiomers as well as individual enantiomers and diastereomers, which arise as a consequence of structural asymmetry in selected compounds of the present series.
The compounds of Formulae I-XI may also be solvated, especially hydrated. Hydration may occur during manufacturing of the compounds or compositions comprising the compounds, or the hydration may occur over time due to the hygroscopic nature of the compounds.
The term xe2x80x9carylxe2x80x9d as employed herein by itself or as part of another group refers to monocyclic or bicyclic aromatic groups containing from 6 to 12 carbons in the ring portion, preferably 6-10 carbons in the ring portion, such as phenyl, naphthyl or tetrahydronaphthyl.
The term xe2x80x9cheteroarylxe2x80x9d as employed herein refers to groups having 5 to 14 ring atoms; 6, 10 or 14 xcfx80 electrons shared in a cyclic array; and containing carbon atoms and 1, 2 or 3 oxygen, nitrogen or sulfur heteroatoms (where examples of heteroaryl groups are: thienyl, benzo[b]thienyl, naphtho[2,3-b]thienyl, thianthrenyl, furyl, pyranyl, isobenzofuranyl, benzoxazolyl, chromenyl, xanthenyl, phenoxathiinyl, 2H-pyrrolyl, pyrrolyl, imidazolyl, pyrazolyl, pyridyl, pyrazinyl, pyrimidinyl, pyridazinyl, indolizinyl, isoindolyl, 3H-indolyl, indolyl, indazolyl, purinyl, 4H-quinolizinyl, isoquinolyl, quinolinyl, tetrahydroquinolinyl, phthalazinyl, naphthyridinyl, quinazolinyl, cinnolinyl, pteridinyl, 4aH-carbazolyl, carbazolyl, xcex2-carbolinyl, phenanthridinyl, acridinyl, perimidinyl, phenanthrolinyl, phenazinyl, isothiazolyl, phenothiazinyl, isoxazolyl, furazanyl and phenoxazinyl groups).
The term xe2x80x9caralkylxe2x80x9d or xe2x80x9carylalkylxe2x80x9d as employed herein by itself or as part of another group refers to C1-6 alkyl groups having an aryl substituent, such as benzyl, phenylethyl or 2-naphthylmethyl.
The term xe2x80x9ccycloalkylxe2x80x9d as employed herein by itself or as part of another group refers to cycloalllyl groups containing 3 to 9 carbon atoms, preferably 4 to 7 carbon atoms. Typical examples are cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl and cyclononyl.
The term xe2x80x9chalogenxe2x80x9d or xe2x80x9chaloxe2x80x9d as employed herein by itself or as part of another group refers to chlorine, bromine, fluorine or iodine with chlorine being preferred.
Scheme Ia illustrates but is not limited to the preparation of compounds of Examples 1, 5, 8, 9, 11, and 12. 
Each of R1 through R3, R6 through R9, n and m is as defmed above; Pa is a hydroxyl protecting group or hydrogen, and Pb is an amino protecting group.
Phenols 1 (where P is H) are converted to monosulfonates 2 by treatment with appropriate sulfonyl chlorides. Preferred conditions include treating phenol 1 with a sulfonyl chloride in a biphasic system composed of ether and an aqueous phase saturated with NaHCO3. Alternatively, the reaction may be effected first by deprotonating 1 with a strong base, most preferably NaH, in a polar organic solvent, such as DMF or tetrahydrofliran, followed by treating the deprotonated phenol with the sulfonyl chloride. Still alternatively, phenol 1, in a typical organic solvent, such as methylene chloride, may be converted to 2 by treating the phenol with sulfonyl chloride in the presence of an amine base, such as N-methylmorpholine.
Phenols 1 may be monoprotected (Pa is a protecting group) with a variety of protecting groups known in the art, such as esters and benzyl ethers (Green, T. W. and Wuts, P. G. M., Protective Groups in Organic Synthesis, 2nd edition, John Wiley and Sons, Inc., New York (1991)). Deprotection of the hydroxyl groups is routinely accomplished using reaction conditions well-known in the art. For example, deprotection of benzyl ethers may be effected through catalytic hydrogenation using palladium on carbon as a catalyst in solvents such as ethanol or tetrahydrofuran. Deprotection of an acetate is accomplished by basic hydrolysis, most preferably with sodium hydroxide in aqueous tetrahydrofuran.
Phenols 2 are coupled to 3 (for Lxe2x95x90OH) using a Mitsunobu coupling procedure (Mitsunobu, O., Synthesis 1 (1981)) to provide 4. Preferred coupling conditions include using a trialkylphosphine or triarylphosphine, such as triphenylphosphine, in a suitable solvent such as tetrahydrofuran or methylene chloride, and a dialkyl azodicarboxylate, such as diethyl azodicarboxylate. In some cases, it is advantageous to add an amine base such as N-methylmorpholine. The amine terminus of 3 is protected with a protecting group Pb that is readily removed from 4. Amino-protecting groups are well known in the art (Greene, T. W. and Wuts, P. G. M., Protective Groups in Organic Synthesis, 2nd edition, John Wiley and Sons, Inc., New York (1991)). Deprotection of the amino group is effected by employing reaction conditions that are well known in the art. For example, the t-butoxycarbonyl (BOC) may be removed by exposure to strongly acidic medium, such as hydrogen chloride, in a suitable solvent, such as dioxane, or a mixed trifluoroacetic acid/methylene chloride solvent system. Benzyloxycarbonyl (CBz) groups may be removed by hydrogen using palladium on carbon as a catalyst in solvents such as ethanol or tetrahydrofuran. The resulting amine is then converted to amidine 5 in a manner similar to the procedure described by Nagahara et. al., J. Med. Chem. 37(8):1200-1207 (1994) wherein the amine is treated with an appropriate imidate in the presence of a base such as N,N-diisopropylethylamine in an appropriate solvent such as DMF. Alternatively, the amine is treated with an appropriate imidate in the presence of a base, such as sodium hydroxide, in an appropriate solvent, such as methanol. Scheme Ib illustrates but is not limited to the preparation of compounds of Examples 2 and 13. 
R1-R3, R6-R8, n, m Pa and Pb are each as defined above.
Aryl ethers 8 are synthesized in a fashion analogous to synthesis of 5. Phenol 1 (P is H) is converted to derivative 6 by treating 1 with a strong base, preferably NaH, in a suitable solvent such as DMF, followed by addition of a reactive alkyl or benzyl compound, R1CH2X (where X is a reactive functional group such as iodide, chloride, bromide or alkylsulfonate). Alternatively, the Mitsunobu Reaction may be used with an appropriate R1CH2X (Xxe2x95x90OH) using the reaction conditions described above. The use of suitable alcohol protecting groups (Pa), such as esters, to suppress over-alkylation, is well known in the art (Greene, T. W. and Wuts, P. G. M., Protective Groups in Organic Synthysis, 2nd edition, John Wiley and Sons, Inc., New York (1991)). The protecting group may then be removed using well-known techniques, for example by hydrolysis with aqueous NaOH, when an ester protecting group is employed. Phenol 6 is then converted to amidine 8 using the conditions described for formation of 5.
Scheme II illustrates, but is not limited to, the preparation of compounds exemplified by Examples 3, 9 and 10. 
R1-R3, R6-R10, n, m, Pa and Pb are as defined above.
According to Scheme II, a nitrophenol 9 may be coupled to compound 3 by standard techniques. Preferably, the reaction is effected by the Mitsunobu reaction (where L is OH). Alternatively, 9 may be treated with a base, such as NaH, in a suitable solvent such as DMF or THF, followed by addition of 3 (where L is a reactive group, such as Cl, Br, I or alkylsulfonate). The nitro group is thereafter reduced, for example, by catalytic reduction using palladium on carbon in a suitable solvent such as ethanol or tetrahydrofuran. The resulting product in then treated with an appropriate sulfonyl chloride (R1SO2Cl) to provide 11. Removal of the amine protecting group Pb is accomplished by techniques known in the art. For example, the t-butoxycarbonyl (BOC) is removed by exposure to a strongly acidic medium, such as hydrogen chloride in a suitable solvent such as dioxane or trfluoroacetic acid in methylene chloride. Benzyloxycarbonyl (CBz) groups are removed by catalytic hydrogen using palladium on carbon as a catalyst in solvents such as ethanol or tetrahydrofuran.
The resulting amine is then converted to amidine 12 in a manner similar to the procedure described by Nagahara et. al., J. Med. Chem. 37(8):1200-1207 (1994) wherein the amine is treated with an appropriate imidate in the presence of a base such as N,N-diisopropylethylamine in an appropriate solvent such as DMF. Alternatively, the amine is treated with an appropriate imidate in the presence of a base such as sodium hydroxide as base in an appropriate solvent such as methanol. N-Substituted sulfonamide derivative 13 is obtained by alkylation of 11 employing a suitable alkylating agent (R10X) in the presence of a base, most preferably Cs2CO3 using a polar solvent such as DMF. Deprotection and amidination are then executed in a manner similar to the conversion of 11 to 12.
Scheme III illustrates but is not limited to the preparation of compounds of Example 4. 
R1-R3, R7-R10, n, m and Pb are each as defined above.
According to Scheme III, nitroaniline 14 is converted to a sulfonamide by treatment with an appropriate sulfonyl chloride R1SO2Cl in the presence of a weak base, such as N-methylmorpholine. The resulting sulfonamide nitrogen is alkylated with a suitable alkylating agent (R10X) in the presence of a base, preferably an alkali metal carbonate such as Cs2CO3 or K2CO3, using a polar solvent, such as DMF, to provide intermediate 15. After reduction of the nitro group, the resulting aniline is coupled to a carboxylic acid, 16, to provide amide 17. Amide coupling may be performed using any of a number of common peptide coupling reagents. Preferably, one of 1,3-dicyclohexylcarbodiimide or Castro""s reagent (BOP) are employed (B. Castro et al., Tetrahedron Lett.:1219 (1975)). Alternatively, 17 may be formed by coupling the aniline with the corresponding acid chloride of acid 16 in the presence of an acid scavenger, such as N-methylmorpholine. Amide 17 is converted to amine 18 by reduction of the amide functionality with an appropriate hydride reagent, preferably borane-THF complex or chlorotrimethylsilane and lithium borohydride. This reaction occurs in a suitable polar solvent, such as THF. Removal of the amine protecting group Pb and formation of the amidine as described in Scheme II provides the desired compound 19. Alternatively the amide nitrogen may be alkylated using a strong base, such as sodium hydride, in a suitable polar solvent such as DMF, followed by treatment with an alkylating agent (R10X) to afford intermediate 20.
Reduction of the amide, as executed in the formation of 18, to give 21 followed by deprotection and amidination as previously described provides the analogous compound 22.
Scheme IV illustrates but is not limited to the preparation of compounds of Examples 6, 7, 14, 15, 16, 17 and 18. 
R1-R3, R6 and n are each as defined above.
Monosulfonates 2 are converted to cyano derivatives 24 by exposing 2 to a base, most preferably sodium hydride in a suitable solvent such as DMF, followed by addition 23, where L is a reactive group such as iodide, chloride, bromide, alkyl sulfonate, or aryl sulfonate. Alternatively, the Mitsunobu Reaction may be used with an appropriate alcohol 23, where Lxe2x95x90OH. The nitrile is submitted to arnidino formation conditions such as those described by Nagahara et. al., J. Med Chem. 37(8):1200-1207 (1994), wherein the nitrile is first exposed to a strong acid, preferably hydrogen chloride, in a suitable alcoholic solvent, preferably methanol or ethanol, which converts the nitrile to an imidate. Following brief isolation, the imidate is treated with an appropriate amine HNR6R6 to effect formation of 25. Similarly, benzamidines 28 are prepared from 2 using appropriate benzonitrile derivatives 26.
It is to be understood that in each of the above-mentioned schemes, an additional substituent, R4, may be present on the phenyl ring of the starting material.
For medicinal use, the pharmaceutically acceptable acid addition salts, those salts in which the anion does not contribute significantly to toxicity or pharmacological activity of the organic cation, are preferred. The acid addition salts are obtained either by reaction of an organic base of Formulae I-XI with an organic or inorganic acid, preferably by contact in solution, or by any of the standard methods detailed in the literature available to any practitioner skilled in the art. Examples of useful organic acids are carboxylic acids such as maleic acid, acetic acid, tartaric acid, propionic acid, fumaric acid, isethionic acid, succinic acid, cyclamic acid, pivalic acid and the like; useful inorganic acids are hydrohalide acids such as HCl, HBr, HI; sulfuric acid; phosphoric acid and the like. Preferred acids for forming acid addition salts include HCl and acetic acid.
The compounds of the present invention represent a novel class of potent inhibitors of metallo, acid, thiol and serine proteases. Examples of the serine proteases inhibited by compounds within the scope of the invention include leukocyte neutrophil elastase, a proteolytic enzyme implicated in the pathogenesis of emphysema; chymotrypsin and trypsin, digestive enzymes; pancreatic elastase, and cathepsin G, a chymotrypsin-like protease also associated with leukocytes; thrombin and factor Xa, proteolytic enzymes in the blood coagulation pathway. Inhibition of thermolysin, a metalloprotease, and pepsin, an acid protease, are also contemplated uses of compounds of the present invention. The compounds of the present invention are preferably employed to inhibit trypsin-like proteases.
An end use application of the compounds that inhibit chymotrypsin and trypsin is in the treatment of pancreatitis. For their end-use application, the potency and other biochemical parameters of the enzyme-inhibiting characteristics of the compounds of the present invention is readily ascertained by standard biochemical techniques well-known in the art. Actual dose ranges for their specific end-use application will, of course, depend upon the nature and severity of the disease state of the patient or animal to be treated, as determined by the attending diagnostician. It is expected that a useful dose range will be about 0.01 to 10 mg per kg per day for an effective therapeutic effect.
Compounds of the present invention that are distinguished by their ability to inhibit either factor Xa or thrombin may be employed for a number of therapeutic purposes. As factor Xa or thrombin inhibitors, compounds of the present invention inhibit thrombin production. Therefore, these compounds are useful for the treatment or prophylaxis of states characterized by abnormal venous or arterial thrombosis involving either thrombin production or action. These states include, but are not limited to, deep vein thrombosis; disseminated intravascular coagulopathy which occurs during septic shock, viral infections and cancer; myocardial infarction; stroke; coronary artery bypass; hip replacement; and thrombus formation resulting from either thrombolytic therapy or percutaneous transluminal coronary angioplasty (PCTA). The compounds of the present invention may also be used as an anticoagulant in extracorporeal blood circuits.
By virtue of the effects of both factor Xa and thrombin on a host of cell types, such as smooth muscle cells, endothelial cells and neutrophils, the compounds of the present invention find additional use in the treatment or prophylaxis of adult respiratory distress syndrome; inflammatory responses, such as edema; reperfusion damage; atherosclerosis; and restenosis following an injury such as balloon angioplasty, atherectomy, and arterial stent placement.
The compounds of the present invention may be useful in treating neoplasia and metastasis as well as neurodegenerative diseases, such as Alzheimer""s disease and Parkinson""s disease.
When employed as thrombin or factor Xa inhibitors, the compounds of the present invention may be administered in an effective amount within the dosage range of about 0.1 to about 500 mg/kg, preferably between 0.1 to 10 mg/kg body weight, on a regimen in single or 2-4 divided daily doses.
When employed as inhibitors of thrombin, the compounds of the present invention may be used in combination with thrombolytic agents such as tissue plasminogen activator, streptokinase, and urokinase. Additionally, the compounds of the present invention may be used in combination with other antithrombotic or anticoagulant drugs such as, but not limited to, fibrinogen antagonists and thromboxane receptor antagonists.
Human leucocyte elastase is released by polymorphonuclear leukocytes at sites of inflammation and thus is a contributing cause for a number of disease states. Thus, compounds of the present invention are expected to have an anti-inflammatory effect useful in the treatment of gout, rheumatoid arthritis and other inflammatory diseases, and in the treatment of emphysema. Cathepsin G has also been implicated in the disease states of arthritis, gout and emphysema, and in addition, glomerulonephritis and lung infestations caused by infections in the lung. In their end-use application the enzyme inhibitory properties of the compounds of Formulae I-XI is readily ascertained by standard biochemical techniques that are well-known in the art.
The neutrophil elastase inhibitory properites of compounds within the scope of the present invention are determined by the following method. Neutrophil elastase is prepared by the procedure described by Baugh et al., Biochemistry 15: 836 (1979). Enzyme assays are conducted substantially according to the procedure disclosed by Nakajima et al., J. Biol. Chem. 254: 4027 (1979), in assay mixtures containing 0.10 M Hepes (N-2-hydroxyethylpiperazine-Nxe2x80x2-2-ethanesulfonic acid) buffer, pH 7.5; 0.5 M NaCl; 10% dimethylsulfoxide; and 1.50xc3x9710xe2x88x924 M MeOSuc-Ala-Ala-Pro-Val-p-nitroanilide as substrate. Inhibitors are evaluated by comparing enzymatic activity measured in the presence and absence of inhibitor.
The Cathepsin G inhibitory properties of compounds within the scope of the present invention are determined by the following method. A preparation of partially purified human Cathepsin G is obtained by the procedure of Baugh et al., Biochemistry 15: 836 (1979). Leukocyte granules are a major source for the preparation of leukocyte elastase and cathepsin G (chymotrypsin-like activity). Leukocytes are lysed and granules are isolated. The leukocyte granules are extracted with 0.20 M sodium acetate, pH 4.0, and extracts are dialyzed against 0.05 M Tris buffer, pH 8.0 containing 0.05 M NaCl overnight at 4xc2x0 C. A protein fraction precipitates during dialysis and is isolated by centrifugation. This fraction contains most of the chymotrypsin-like activity of leukocyte granules. Specific substrates are prepared for each enzyme, namely MeOSuc-Ala-Ala-Pro-Val-p-nitroanilide and Suc-Ala-Ala-Pro-Phe-p-nitroanilide. The latter is not hydrolyzed by leukocyte elastase. Enzyme preparations are assayed in 2.00 mL of 0.10 M Hepes buffer, pH 7.5, containing 0.50 M NaCl, 10% dimethylsulfoxide and 0.0020 M Suc-Ala-Ala-Pro-Phe-pnitroanilide as a substrate. Hydrolysis of the p-nitroanilide substrate is monitored at 405 nm and at 25xc2x0 C.
Useful dose range for the application of compounds of the present invention as neutrophil elastase inhibitors and as Cathepsin G inhibitors will of course depend upon the nature and severity of the disease state, as determined by the attending diagnostician, with the range of 0.01 to 10 mg/kg of body weight, per day, being useful for the aforementioned disease states.
Compounds of the present invention that inhibit urokinase or plasminogen activator are potentially useful in treating excessive cell growth disease state. As such the compounds of the present invention may also be useful in the treatment of benign prostatic hypertrophy and prostatic carcinoma, the treatment of psoriasis, and in their use as abortifacients. For their end-use application, the potency and other biochemical parameters of the enzyme inhibiting characteristics of the compounds of the present invention are readily ascertained by standard biochemical techniques well-known in the art. Actual dose ranges for their specific end-use application will, of course, depend upon the nature and severity of the disease state of the patient or animal to be treated as determined by the attending diagnostician. It is to be expected that the general end-use application dose range will be about 0.01 to 10 mg per kg per day for an effective therapeutic effect.
Additional uses for compounds of the present invention include analysis of commercial reagent enzymes for active site concentration. For example, chymotrypsin is supplied as a standard reagent for use in clinical quantitation of chymotrypsin activity in pancreatic juices and feces. Such assays are diagnostic for gastrointestinal and pancreatic disorders. Pancreatic elastase is also supplied commercially as a reagent for quantitation of xcex11-antitrypsin in plasma. Plasma xcex11-antitrypsin increases in concentration during the course of several inflammatory diseases, and xcex11-antitrypsin deficiencies are associated with increased incidence of lung disease. Compounds of the present invention can be used to enhance the accuracy and reproducibility of this assay by titrametric standardization of the commercial elastase supplied as a reagent. See, U.S. Pat. No. 4,499,082.
Protease activity in certain protein extracts during purification of particular proteins is a recurring problem which can complicate and compromise the results of protein isolation procedures. Certain proteases present in such extracts can be inhibited during purification steps by compounds of the present invention, which bind tightly to various proteolytic enzymes.
The pharmaceutical compositions of the invention can be administered to any animal that can experience the beneficial effects of the compounds of the invention. Foremost among such animals are humans, although the invention is not intended to be so limited.
The pharmaceutical compositions of the present invention can be administered by any means that achieve their intended purpose. For example, administration can be by parenteral, subcutaneous, intravenous, intramuscular, intraperitoneal, transdermal, buccal, or ocular routes. Alternatively, or concurrently, administration can be by the oral route. The dosage administered will be dependent upon the age, health, and weight of the recipient, kind of concurrent treatment, if any, frequency of treatment, and the nature of the effect desired.
In addition to the pharmacologically active compounds, the new pharmaceutical preparations can contain suitable pharmaceutically acceptable carriers comprising excipients and auxiliaries that facilitate processing of the active compounds into preparations that can be used pharmaceutically.
The pharmaceutical preparations of the present invention are manufactured in a manner that is, itself, known, for example, by means of conventional mixing, granulating, dragee-making, dissolving, or lyophilizing processes. Thus, pharmaceutical preparations for oral use can be obtained by combining the active compounds with solid excipients, optionally grinding the resulting mixture and processing the mixture of granules, after adding suitable auxiliaries, if desired or necessary, to obtain tablets or dragee cores.
Suitable excipients are, in particular, fillers such as saccharides, for example, lactose or sucrose, mannitol or sorbitol, cellulose preparations and/or calcium phosphates, for example, tricalcium phosphate or calcium hydrogen phosphate, as well as binders, such as, starch paste, using, for example, maize starch, wheat starch, rice starch, potato starch, gelatin, tragacanth, methyl cellulose, hydroxypropylmethylcellulose, sodium carboxymethylcellulose, and/or polyvinyl pyrrolidone. If desired, disintegrating agents can be added, such as, the above-mentioned starches and also carboxymethyl-starch, cross-linked polyvinyl pyrrolidone, agar, or alginic acid or a salt thereof, such as, sodium alginate. Auxiliaries are, above all, flow-regulating agents and lubricants, for example, silica, talc, stearic acid or salts thereof, such as, magnesium stearate or calcium stearate, and/or polyethylene glycol. Dragee cores are provided with suitable coatings that, if desired, are resistant to gastric juices. For this purpose, concentrated saccharide solutions can be used, which may optionally contain gum arabic, talc, polyvinyl pyrrolidone, polyethylene glycol, and/or titanium dioxide, lacquer solutions and suitable organic solvents or solvent mixtures. In order to produce coatings resistant to gastric juices, solutions of suitable cellulose preparations, such as, acetylcellulose phthalate or hydroxypropylmethyl-cellulose phthalate, are used. Dye stuffs or pigments can be added to the tablets or dragee coatings, for example, for identification or in order to characterize combinations of active compound doses.
Other pharmaceutical preparations which can be used orally include push-fit capsules made of gelatin, as well as soft, sealed capsules made of gelatin and a plasticizer, such as, glycerol or sorbitol. The push-fit capsules can contain the active compounds in the form of granules that may be mixed with fillers such as lactose, binders such as starches, and/or lubricants such as talc or magnesium stearate and, optionally, stabilizers. In soft capsules, the active compounds are preferably dissolved or suspended in suitable liquids, such as, fatty oils or liquid paraffin. In addition, stabilizers may be added.
Suitable formulations for parenteral administration include aqueous solutions of the active compounds in water-soluble form, for example, water-soluble salts, alkaline solutions and cyclodextrin inclusion complexes. Especially preferred alkaline salts are ammonium salts prepared, for example, with Tris, choline hydroxide, Bis-Tris propane, N-methylglucamine, or arginine. One or more modified or unmodified cyclodextrins can be employed to stabilize and increase the water solubility of compounds of the present invention. Useful cyclodextrins for this purpose are disclosed in U.S. Pat. Nos. 4,727,064, 4,764,604, and 5,024,998.
In addition, suspensions of the active compounds as appropriate oily injection suspensions can be administered. Suitable lipophilic solvents or vehicles include fatty oils, for example, sesame oil, or synthetic fatty acid esters, for example, ethyl oleate or triglycerides or polyethylene glycol-400 (the compounds are soluble in PEG-400). Aqueous injection suspensions can contain substances that increase the viscosity of the suspension, for example, sodium carboxymethyl cellulose, sorbitol, and/or dextran. Optionally, the suspension may also contain stabilizers.